标签：Calcein-AM 与 Calcein 相比具有增强的疏水性

Dojindo,Cellstain钙黄绿素AM溶液/1/C396,Calcein-AM 与 Calcein 相比具有增强的疏水性

Calcein-AM 与 Calcein 相比具有增强的疏水性，因此很容易通过活细胞的细胞膜。 钙黄绿素-AM 渗入细胞质后，被酯酶水解成钙黄绿素，钙黄绿素留在细胞内（图 1）。 在其他试剂中，包括 BCECF-AM 和羧基荧光素二乙酸酯，Calcein-AM 是最适合用于染色活细胞的荧光探针，因为它的细胞毒性低。 钙黄绿素不抑制任何细胞功能，例如淋巴细胞的增殖或趋化性。 此外，使用钙黄绿素的活力测定是可靠的，并且与标准的 51Cr 释放测定具有良好的相关性。 calcein 的激发和发射波长分别为 490 nm 和 515 nm

Fig. 1 Cell staining mechanism

Staining Procedure1.Prepare 1 mM Calcein-AM solution with DMSO and dilute to prepare 1-50 μM Calcein-AM solution with PBS.a)2.Add Calcein-AM solution with 1/10 of the volume of cell culture medium to the cell culture.b)3.Incubate the cell at 37ºC for 15-30 min.4.Wash cells twice with PBS or an appropriate buffer.5.Observe the cells under a fluorescence microscope with 490 nm excitation and 515 nm emission filters.

a) If the Calcein-AM has difficulty loading into cells, use a detergent such as Pluronic F127.b) Or you may replace the culture medium with 1/10 concentration of Calcein-AM buffer solution.

1. K. McGinnes, et al., A Fluorescence NK Assay Using Flow Cytometry. J Immunol Methods. 1986;86:7-15.2. S. J. Morris, Real-time Multi-wavelength Fluorescence Imaging of Living Cells. BioTechniques. 1990;8:296-308.3. S. A. Weston, et al., New Fluorescent Dyes for Lymphocyte Migration Studies Analysis by Flow Cytometry and Fluorescent Microscopy. J Immunol Methods. 1990;133:87-97.4. D. M. Callewaert, et al., Characterization of Effector-Target Conjugates for Cloned Human Natural Killer and Human Lymphokine Activated Killer Cells by Flow Cytometry. Cytometry. 1991;12:666-676.5. H. Xie, et al., Intercellular Communication Through Gap Junctions Is Reduced in Senescent Cell. Biophys J. 1992;62:45-47.6. S. A. Weston, et al., Calcein: a Novel Marker for Lymphocytes Which Enter Lymph Nodes. Cytometry. 1992;13:739-749.7. X. M. Wang, et al., A New Microcellular Cytotoxicity Test Based on Calcein AM Release. Hum Immunol. 1993;37:264-270.8. N. G. Papadopoulos, et al., An Improved Fluorescence Assay for the Determination of Lymphocyte-Mediated Cytotoxicity Using Flow Cytometry. J Immunol Methods. 1994;177:101-111. 9. L. S. D. Clerck, et al., Use of Fluorescent Dyes in the Determination of Adherence of Human Leucocytes to Endothelial Cells and the Effects of Fluorochromes on Cellular Function. J Immunol Methods. 1994;172:115-124.10. H. Ohata, et al., Confocal Imaging Analysis of ATP-Induced Ca2+ Response in Individual Endothelial Cells of the Artery in Situ. Am J Physiol. 1997;272:C1980-C1987.

Dojindo,Cellstain-Calcein AM/1/C326,Calcein-AM 与 Calcein 相比具有增强的疏水性

Calcein-AM 与 Calcein 相比具有增强的疏水性，因此很容易通过活细胞的细胞膜。 钙黄绿素-AM 渗入细胞质后，被酯酶水解成钙黄绿素，钙黄绿素留在细胞内（图 1）。 在其他试剂中，包括 BCECF-AM 和羧基荧光素二乙酸酯，Calcein-AM 是最适合用于染色活细胞的荧光探针，因为它的细胞毒性低。 钙黄绿素不抑制任何细胞功能，例如淋巴细胞的增殖或趋化性。 此外，使用钙黄绿素的活力测定是可靠的，并且与标准的 51Cr 释放测定具有良好的相关性。 钙黄绿素的激发和发射波长分别为 490 nm 和 515 nm。

Fig. 1 Cell staining mechanism

*caution* Please tap the tube before opening, and open it with care. The content may have relocated from the bottom of the tube during the shipping.

*caution*Please tap the tube before opening, and open it with care. The content may have relocated from the bottom of the tube during the shipping.

Staining Procedure1.Prepare 1 mM Calcein-AM solution with DMSO and dilute to prepare 1-50 μM Calcein-AM solution with PBS.a) 2.Add Calcein-AM solution with 1/10 of the volume of cell culture medium to the cell culture.b) 3.Incubate the cell at 37ºC for 15-30 min. 4.Wash cells twice with PBS or an appropriate buffer. 5.Observe the cells under a fluorescence microscope with 490 nm excitation and 515 nm emission filters.

a) If the Calcein-AM has difficulty loading into cells, use a detergent such as Pluronic F127.b) Or you may replace the culture medium with 1/10 concentration of Calcein-AM buffer solution.

1. K. McGinnes, et al., A Fluorescence NK Assay Using Flow Cytometry. J Immunol Methods. 1986;86:7-15.2. S. J. Morris, Real-time Multi-wavelength Fluorescence Imaging of Living Cells. BioTechniques. 1990;8:296-308.3. S. A. Weston, et al., New Fluorescent Dyes for Lymphocyte Migration Studies Analysis by Flow Cytometry and Fluorescent Microscopy. J Immunol Methods. 1990;133:87-97.4. D. M. Callewaert, et al., Characterization of Effector-Target Conjugates for Cloned Human Natural Killer and Human Lymphokine Activated Killer Cells by Flow Cytometry. Cytometry. 1991;12:666-676.5. H. Xie, et al., Intercellular Communication Through Gap Junctions Is Reduced in Senescent Cell. Biophys J. 1992;62:45-47.6. S. A. Weston, et al., Calcein: a Novel Marker for Lymphocytes Which Enter Lymph Nodes. Cytometry. 1992;13:739-749.7. X. M. Wang, et al., A New Microcellular Cytotoxicity Test Based on Calcein AM Release. Hum Immunol. 1993;37:264-270.8. N. G. Papadopoulos, et al., An Improved Fluorescence Assay for the Determination of Lymphocyte-Mediated Cytotoxicity Using Flow Cytometry. J Immunol Methods. 1994;177:101-111.9. L. S. D. Clerck, et al., Use of Fluorescent Dyes in the Determination of Adherence of Human Leucocytes to Endothelial Cells and the Effects of Fluorochromes on Cellular Function. J Immunol Methods. 1994;172:115-124.10. H. Ohata, et al., Confocal Imaging Analysis of ATP-Induced Ca2+ Response in Individual Endothelial Cells of the Artery in Situ. Am J Physiol. 1997;272:C1980-C1987.